Sustained release bimatoprost, bimatoprost analogs, prostamides and prostaglandins for fat reduction

ABSTRACT

The present invention is directed to compositions and methods for injection into fat deposits for sustained release of compounds which result in localized fat reduction.

CROSS REFERENCE TO RELATED APPLICATION

This application claims the benefit of U.S. Provisional PatentApplication Ser. No. 61/811,682, filed Apr. 12, 2013, the entiredisclosure of which is incorporated herein by reference.

SUMMARY OF THE INVENTION

The present invention is directed to compositions and methods for thesustained release of bimatoprost, bimatoprost analogs, bimatoprostprodrugs, prostamides, prostaglandins, prostaglandin analogs andprostaglandin derivatives from injectable and implantable depots for thepurpose of fat reduction including localized fat reduction.

Topical bimatoprost has been shown to effectively prevent apidocyteformation and maturation and to atrophy adipocytes in animal modelsafter topical administration. Furthermore, clinical evidence of fatreduction after topical administration of bimatoprost has been reported.The present invention is directed to sustained release methods andformulations of bimatoprost, bimatoprost analogs, bimatoprost prodrugs,prostamides, prostaglandins, prostaglandin analogs and derivatives andprostaglandin analogs such as latanoprost and travoprost for localizedfat reduction.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows in vitro release profiles of brimatoprost with injectablePLGA implants;

FIG. 2 shows in vitro release profiles of brimatoprost with injectableSynBiosys implants;

FIGS. 3A and 3B show 100 day ReGel 100 or ReGel B release data;

FIGS. 4A-4C show a comparison of latanoprost release data of ReGel 100or ReGel B delivery systems;

FIG. 5 shows a 2-D MR image 40 minutes post-injection orientedlongitudinal through the gastrocnemius muscle to show the MGL and MGMmuscle groups in each leg. The right leg was injected withHA/Albumin-Gadolinium, the left leg with Albumin-Gadolinium alone;

FIG. 6A shows the release rate of bimatoprost in a formulation of 20%Bimatoprost, 45% R203s, 20% RG752s 10% R202H, 5% PEG-3350;

FIG. 6B shows the release rate of bimatoprost in a formulation shown atthe bottom of FIG. 6B;

FIG. 6C shows the release rate of Compound #1 of the formulations ofTable I;

FIG. 7A shows bimatoprost microspheres which can be used for sustainedrelease of bimatoprost for localized fat reduction;

FIG. 7B shows the release rate of bimatoprost from the latanoprostmicrospheres;

FIGS. 8A and 8B show shows 10% bimatoprost in diethyl glycol dibenzoate(gel); and,

FIG. 8C shows an example of bimatoprost release from a 10% Bimatoprostin Diethyl Glycol Dibenzoate depot (gel).

Some embodiments of the invention are included in the followingparagraphs:

1) A method of fat reduction comprising injecting a sustained releaseformulation of a compound selected from the group consisting ofbimatoprost, bimatoprost analogs, bimatoprost prodrugs, prostamides,prostaglandins, prostaglandin analogs, latanoprost and travoprost andprostaglandin derivatives and mixtures thereof into a fat deposit.

2) The method of paragraph 1 wherein the compound is selected from thegroup consisting of bimatoprost, latanoprost, travoprost and Compound #1and mixtures thereof.

3) The method of paragraph 1 wherein the sustained release formulationis selected from the group consisting of injectable depots, gelsuspensions, a ReGel delivery system, a hyaluronic acid releaseplatform, implants, microspheres, macrospheres and injectable solvents.

4) The method of paragraphs 2 or 3 wherein the compound is bimatoprost.

5) The method of paragraphs 1-4 wherein the sustained releaseformulation is injected directly into the fat deposit.

6) The method of paragraph 1 wherein the fat reduction is localized fatreduction at and around the injection site.

7) The method of paragraphs 1 or 3 wherein the sustained releaseformulation is an implant with the formulation of about 20% bimatoprost,about 45% R203s, about 20% RG752s, about 10% R202H and about 5%PEG-3350.

8) The method of paragraphs 1 or 3 wherein the method results in atrophyof both brown and white adipocytes and results in localized fatreduction.

9) The method of paragraph 7 wherein the implant releases bimatoprost inthe fat deposit for over a period of 100 days.

10) The method of paragraph 1 wherein the sustained release formulationreleases the compound systemically to target a fat deposit at a locationin the body that is not at the location of the sustained releasedformulation or at difficult to reach areas.

11) The method of paragraphs 1 or 10 wherein the sustained releaseformulations are injected or implanted at a location that permitsreduction of abdominal fat deposits, visceral fat deposits, epicardialfat deposits, subcutaneous fat deposits and ectopic fat deposits asnon-limiting examples.

12) A composition for use in localized fat reduction wherein thecomposition is a sustained release composition selected from the groupconsisting of injectable depots, gel suspensions, a ReGel deliverysystem, a hyaluronic acid based platform, implants, microspheres,macrospheres and injectable solvents.

13) The composition of paragraph 12 wherein the composition furthercomprises a compound selected from the group consisting of bimatoprost,bimatoprost analogs, bimatoprost prodrugs, prostamides, prostaglandins,prostaglandin analogs, latanoprost and travoprost and prostaglandinderivatives.

14) The composition of paragraph 12 and 13 wherein the sustained releasecomposition is a Regel delivery system and the compound is bimatoprost.

15) The composition of paragraph 14 wherein the composition is injectedinto a localized fat deposit.

16) The composition of paragraph 15 wherein the composition is injectedat multiple injection sites into a single localized fat deposit.

17) The composition of paragraph 15 wherein the composition releasesbimatoprost into the localized fat deposit for over a period of 100days.

18) The composition of paragraph 12 or 13 wherein the sustained releaseformulation is an implant comprised of about 20% bimatoprost, about 45%R203s, about 20% RG752s, about 10% R202H and about 5% PEG-3350.

19) The composition of paragraph 18 wherein the composition is injectedinto at least one selected from the group consisting of abdominal fatdeposits, visceral fat deposits, epicardial fat deposits, subcutaneousfat deposits and ectopic fat deposits.

20) The composition of paragraph 18 wherein injection of the compositionresults in atrophy of adipocytes in the localized fat deposit andreduction of localized fat.

21) The composition of paragraph 12 or 13 wherein the sustained releaseformulation is an implant comprised of at least one polymer selectedfrom the group consisting of poly(d,l-lactide-co-glycolide), poly(d,l-lactide), poly(caprolactone), poly(dioxanone), poly(ethyleneglycol), poly(ortho-ester), polyesters, poly(phosphazine), poly(phosphate ester), polycaprolactone, silicone, natural polymers such aslatex, gelatin or collagen, or polymeric blends and the compound isselected from the group consisting of bimatoprost, latanoprost,travoprost and mixtures thereof.

22) The composition of paragraph 12 or 13 wherein the sustained releaseformulation is a gel suspension comprised of at least one compoundselected from the group consisting of sodium hyaluronate, crosslinkedhyaluronic acid, chondroitin sulfate, cellulosics, gelatin, collagen,glycosaminoclycans, or other synthetic or naturally occurringpolysaccharides and the compound is selected from the group consistingbimatoprost, latanoprost, travoprost and mixtures thereof.

23) The composition of paragraph 22 wherein the gel suspension is athermal gelling delivery system.

24) The composition of paragraph 22 wherein the thermal gelling systemis comprised of solutions of A-B-A or B-A-B triblock copolymers or B-Ablock copolymers.

25) The composition of paragraph 12 or 13 wherein the sustained releaseformulation is an injectable depot with biocompatible solvents selectedfrom the group consisting of DMSO, NMP and DMAC or mixtures thereof.

DETAILED DESCRIPTION OF THE INVENTION

Bimatoprost and other compounds can be dissolved or dispersed in a gel,in a biodegradable solid implant, or biocompatible solvents containingsolvated polymers, which can form solid depots upon injection.Additionally, thermal gelling delivery systems of bimatoprost may alsobe utilized. Solid implants for sustained release may be comprised ofpoly(d,l-lactide-co-glycolide), poly (d,l-lactide), poly(caprolactone),poly(dioxanone), poly(ethylene glycol), poly(ortho-ester), polyesters,poly(phosphazine), poly(phosphate ester), polycaprolactone, silicone,natural polymers such as latex, gelatin or collagen, or polymericblends. Gel suspensions could contain sodium hyaluronate, crosslinkedhyaluronic acid, chondroitin sulfate, cellulosics, gelatin, collagen,glycosaminoclycans, or other synthetic or naturally occurringpolysaccharides. Biocompatible solvents for injection of in situ formingdepots include DMSO (dimethyl sulfoxide), NMP (N-methylpyrrolidone),DMAC (dimethylacetamide), or other non-aqueous solvents for injection.

Bimatoprost delivery systems and delivery systems for other compoundscan be administered for reduction of adipose tissue through theinjection or implantation of implants or injectable depots. Suchdelivery systems may be used for reduction of local adipose tissue, e.gsubcutaneous fat, and/or as a method for sustained systemic delivery toachieve reduction of visceral fat and other fat pad depositions that arenot easily reached by local administration of the implant or injectionsuch as pericardial fat depositions. Bimatoprost is a low meltingcompound and the ability to sustain its release from multiple deliveryplatforms is surprising. Specific delivery platforms include but are notlimited to injectable bimatoprost delivery depots, in situ formingbimatoprost depots, hyaluronic acid depots, solid form bimatoprostimplants, bimatoprost microspheres and injectable solvent depots.

The delivery systems of the present invention can be injected orimplanted at a location to achieve reduction of subcutaneous fatdeposits and adipose tissue such as abdominal fat, visceral fat,epicardial fat, submental fat, periorbital fat and ectopic fat pads.

EXAMPLE I Injectable Depots

PLGA and multiblock polymers have been shown to release bimatoprost upondepot formation. The polymers and drug are dissolved in a biocompatiblesolvent for both, such as N-methypyrrolidinone, di-methyl acetamide orDMSO. The formulation is sterile filtered, autoclaved, or irradiated forsterility. The solution is filled into a sterile vial or a unit dosesyringe. After injection, the biocompatible solvent diffuses away fromthe depot, leaving behind a firm prostamide or prostaglandin loadedimplant. The depot releases bimatoprost, prostamide or prostaglandin fordays, weeks, or months, as the polymer bioerodes. Drug loading insolution could range from 0.1% to 50%. Polymer loading in solution couldrange from 15% to 50%. Excipients could include poly(ethylene glycol),short chain fatty acids, waxes, cholesterol, aliphatic alcohols,co-solvents, or other compounds which would adjust the hydrophobicity ofthe depot.

With both PLGA and SynBiosys bimatoprost containing injectable depots,the drug was continuously released for at least one month as shown inFIGS. 1 and 2. It is possible to further optimize the release kineticsby varying drug load, polymer concentration, polymer properties,formulation excipients or DMSO volume used for implant preparation.

EXAMPLE II ReGel Delivery System

Polymer systems that undergo phase transitions in response to variousstimuli can also be used. This phase transition results in a significantvolume and or viscosity change in the system. The system can respond topH, ionic environment, temperature, biologic triggers as well as otherchemical and physical triggers. The system comprises one or morepolymers capable of interacting to cause a phase-transition resulting inthe volume or viscosity increases. Examples of polymers includepolyacrylic acid and polyethylene oxide copolymers. Other components ofthe system include excipients known to those experienced in the art.

The system has the further advantage of offering controlled andsustained release of therapeutically active agents to local tissues. Thedrug may be physically entrapped or chemically bound via covalentlinkages, hydrogen binding, ionic interactions, van der Waals forces orhydrophobic interactions. Release of the drug can be controlled byphysical entrapment of the active compound in the transitioned gel.Compounds can also be physically or chemically bound to the polymerscomprising the phase transition gel. The phase transition of the gelserves to create a depot for drug delivery.

A specific example of this invention teaches the use of thermal gellingbimatoprost deliver delivery systems comprised of solutions of A-B-A orB-A-B triblock copolymers or B-A block copolymers whereA=polylactide-co-glycolide (PLGA/PLA) and B=polyethylene oxide (PEO) andlatanoprost. These polymers make up the Regel in situ gelling deliverysystem. Its aqueous solutions have shown to have sol-to-gel transitionbehavior as temperature increases. For drug delivery applications,gelation at physiologically relevant temperature (e.g., 37° C.) isparticularly important and forms the basis for the utility of thesystems for medical and drug delivery purposes.

In the specific example, latanoprost was loaded at 3% loading into ReGel100 or ReGel B i.e. 3 mg drug in 100 ul gel. The system displayedsustained release after thermal gelation with no burst of latanoprost.This is very surprising given the relative low melting point andsolubility of latanoprost. i.e., slow release, no burst. The gelremained for longer than 100 days as shown in FIGS. 3A and 3B.Additional modifications can be made by adding other polymers to thesystem, e.g., CMC, agarose and starch.

EXAMPLE III Hyaluronic Acids

Crosslinked hyaluronic acid has been shown to localize upon injectionproviding a potential sustained release platform. Drug can either beincorporated into the crosslinked hyaluronic acid or conjugated to thevehicle for sustained release. In the case of the former, release anderosion of the platform can be controlled by porosity of the gel, lengthof the crosslinkers and crosslinking density. Alternatively, in thelatter case, bimatoprost or a prostamide analog can be covalently orionically bonded to the hyaluronic backbone through one of severallinkers known to the art. Finally, drug may be incorporated into anothersustained release modality, such as microspheres, then incorporated intothe hyaluronic acid (crosslinked or non-crosslinked) and injected as adelivery platform.

FIG. 5 shows a 2-D MR image 40 minutes post-injection orientedlongitudinal through the gastrocnemius muscle to show the MGL(tripennate gastrocnemius lateralis) and MGM (unipennate gastrocnemiusmedialis) muscle groups in each leg. The right leg was injected withHA/Albumin-Gadolinium, the left leg with Albumin-Gadolinium alone. Theleft leg shows diffuse spread of the Albumin-Gadolinium (blue color)throughout the MGL muscle and crossover to the adjacent MGM muscle. Thisdata shows that cross-linked HA depots can be localized and provide aplatform for the local sustained release of a prostaglandin orprostamide for fat reduction.

EXAMPLE IV Implants

Bimatoprost has been formulated into implants that can be injected orimplanted subcutaneously, into visceral fat or in direct apposition toan organ. An example is the following formulation : 20% Bimatoprost, 45%R203s, 20% RG752s 10% R202H, 5% PEG-3350 and FIG. 6A shows the releaserate of bimatoprost from this formulation (R203S is an ester end-cappedPLA, R202H is an acid end group PLA, RG752S is a 75:25 PLGA with anester end group and PEG_(—)3350 is polyethylene glycol with a molecularweight of 3350). Other implant formulations and their release rates areshown in FIG. 6B.

Another compound (Compound #1) which may be useful for fat reduction isdisclosed below:

Implant formulations with Compound #1 and their properties are in TableI below:

in vitro estimated release release implant implant rate durationExamples Lot # compositions dimension wt (μg) (μg/d) (month) 1 10524-1018.0% API. 92.0% 150 μm × 1.5 mm 36 29 3 R202H 2 10810-061 8.0% API,92.0% 200 μm × 1.5 mm 64 26 6 R203H 3 10810-080 8.0% API, 51.7% 200 μm ×1.5 mm 64 34 4-5 R203S, 23.0% RG752S, 11.5% R202H, 5.8% hexadecanol 410810-116 8.0% API, 18.4% 200 μm × 1.5 mm 64 28 6 R203S, 73.6% R203H

EXAMPLE V Microspheres

Bimatoprost and latanoprost can also be sustained through the use ofPLGA microspheres and macrospheres as shown in FIGS. 7A-7B forlatanoprost. Latanoprost microspheres were manufactured from the PLA andPLGA polymers as shown in the table below. The microspheres weremanufactured by dissolving 20 mg of latanoprost and 100 mg polymer in0.8 ml ethyl acetate. A minimum amount of dichloromethane may be addedto complete dissolution of the polymer. This solution is added to 40 mL1% polyvinyl alcohol aqueous solution via a micro-pipette while mixingat high sheer, 3000 rpm, for 5 minutes with a homogenizer.

After shearing, a milky white emulsion is formed, and it is mildlyagitated in a fume hood for 3-5 hours to allow solvent evaporation. Thisdispersion is then centrifuged at 2000 rpm for 15 min to removesupernatant, and then 10 mL water is added to reconstitute themicrospheres. The final reconstituted micropsheres are lyophilized. Therelease of latanoprost from the microspheres into isotonic phosphatebuffered saline is shown in FIG. 7B

drug entrap PS before freeze drying, Lot load efficiency um numberPolymer % % d10 d90 Mean MP-5 203H 13.5% 81.0% 15.4 59.2 31.9 MP-8 R203S12.2% 73.3% 15.8 64.5 34.7 MP-11 RG755 11.9% 71.3% 17.4 66.3 35.6

EXAMPLE VI Injectable Solvents

Other excipients such as sucrose acetate isobutyrate, ethyl benzoate,benzyl benzoate. tripropionin, diethyl Gglycol dibenzoate among otherscan be used for direct injection subcutaneously or into the fat. FIGS.8A-8B shows 10% bimatoprost in Diethyl Glycol Dibenzoate (gel) and FIG.8C shows an example of bimatoprost release from 10% Bimatoprost inDiethyl Glycol Dibenzoate (gel).

1) A method of fat reduction comprising injecting a sustained releaseformulation of a compound selected from the group consisting ofbimatoprost, bimatoprost analogs, bimatoprost prodrugs, prostamides,prostaglandins, prostaglandin analogs, latanoprost and travoprost andprostaglandin derivatives and mixtures thereof into a fat deposit. 2)The method of claim 1, wherein the compound is selected from the groupconsisting of bimatoprost, latanoprost, travoprost and Compound #1 andmixtures thereof. 3) The method of claim 1, wherein the sustainedrelease formulation is selected from the group consisting of injectabledepots, gel suspensions, a ReGel delivery system, a hyaluronic acidrelease platform, implants, microspheres, macrospheres and injectablesolvents. 4) The method of claim 3, wherein the compound is bimatoprost.5) The method of claim 1, wherein the sustained release formulation isinjected directly into the fat deposit. 6) The method of claim 1,wherein the fat reduction is localized fat reduction at and around theinjection site. 7) The method of claim 3, wherein the sustained releaseformulation is an implant with the formulation of about 20% bimatoprost,about 45% R203s, about 20% RG752s, about 10% R202H and about 5%PEG-3350. 8) The method of claim 1, wherein the method results inatrophy of both brown and white adipocytes and results in localized fatreduction. 9) The method of claim 7, wherein the implant releasesbimatoprost in the fat deposit for over a period of 100 days. 10) Themethod of claim 1, wherein the sustained release formulation releasesthe compound systemically to target a fat deposit at a location in thebody that is not at the location of the sustained release formulation.11) The method of claim 10, wherein the sustained release formulationsare injected or implanted at a location that permits reduction ofabdominal fat deposits, visceral fat deposits, epicardial fat deposits,subcutaneous fat deposits and ectopic fat deposits. 12) A compositionfor use in localized fat reduction wherein the composition is asustained release composition selected from the group consisting ofinjectable depots, gel suspensions, a ReGel delivery system, ahyaluronic acid based platform, implants, microspheres, macrospheres andinjectable solvents. 13) The composition of claim 12, wherein thecomposition further comprises a compound selected from the groupconsisting of bimatoprost, bimatoprost analogs, bimatoprost prodrugs,prostamides, prostaglandins, prostaglandin analogs, latanoprost andtravoprost and prostaglandin derivatives. 14) The composition of claim12, wherein the sustained release composition is a Regel delivery systemand the compound is bimatoprost. 15) The composition of claim 14,wherein the composition is injected into a localized fat deposit. 16)The composition of claim 15, wherein the composition is injected atmultiple injection sites into a single localized fat deposit. 17) Thecomposition of claim 15, wherein the composition releases bimatoprostinto the localized fat deposit for over a period of 100 days. 18) Thecomposition of claim 13, wherein the sustained release formulation is animplant comprised of about 20% bimatoprost, about 45% R203s, about 20%RG752s, about 10% R202H and about 5% PEG-3350. 19) The composition ofclaim 18, wherein the composition is injected into at least one selectedfrom the group consisting of abdominal fat deposits, visceral fatdeposits, epicardial fat deposits, subcutaneous fat deposits and ectopicfat deposits. 20) The composition of claim 18, wherein injection of thecomposition results in atrophy of adipocytes in the localized fatdeposit and reduction of localized fat.